A phenotypic culture system for the molecular analysis of CNS myelination in the spinal cord

Abstract

Studies of central nervous system myelination lack defined in vitro models which would effectively dissect molecular mechanisms of myelination that contain cells of the correct phenotype. Here we describe a co-culture of purified motoneurons and oligodendrocyte progenitor cells, isolated from rat embryonic spinal cord using a combination of immunopanning techniques. This model illustrates differentiation of oligodendrocyte progenitors into fully functional mature oligodendrocytes that myelinate axons. It also illustrates a contribution of axons to the rate of oligodendrocyte maturation and myelin gene expression. The defined conditions used allow molecular analysis of distinct stages of myelination and precise manipulation of inductive cues affecting axonal–oligodendrocyte interactions. This phenotypic in vitro myelination model can provide valuable insight into our understanding of demyelinating disorders, such as multiple sclerosis and traumatic diseases such as spinal cord injury where demyelination represents a contributing factor to the pathology of the disorder.

Recent Advances in Body-on-a-Chip Systems

“…15 However, these strategies are only partial solutions since it will be more difficult with increasing number of organs, and obviously, the development of a universal as well as chemically well-defined media that can support growth and differentiation of all cell types would be a more complete solution. Hickman et al demonstrated using a serum-free medium for supporting coculture of muscle cells derived from human stem cells and sensory neurons, 127 and coculture of motoneurons and oligodendrocyte, 128 as well as in their 4-organ systems, 25 which show great promise for a coculture systems with a larger number of organs. 25 Second, development of biomaterials that meet specifications of different cell types is also needed to further improve current BOC models.…”

Multiorgan Microphysiological Systems for Drug Development: Strategies, Advances, and Challenges

“…[166] The formula were further adapted to support reliable long-term culture of skeletal myotubes, [167] cardiomyocytes, [125] and motoneurons, [22,168] as well as dual culture of motoneurons with sensory neurons to build the stretch reflex arc, [169–171] or with oligodendrocyte progenitor cells to model myelination. [172] This laid a foundation for introducing serum-free cell culture media into the multi-organ systems. Recently, Oleaga et al demonstrated in a four-organ MOM, in which serum-free culture medium maintained hepatic function, cardiac and neuronal electrical activity, and cardiac and skeletal muscle contraction capacity.…”

Self-contained, low-cost Body-on-a-Chip systems for drug development

“…Models for both CNS and PNS myelination have been established to promote neuron-neuron and neuron-muscle communication and to study diseases involving demyelination and injury repair. 26,81,82 A higher degree of neuronal communication is the reflex arc circuit (CNS-PNS link). The sensory and the effector portions have been successfully reproduced in vitro under controlled conditions.…”

“…In 1995, the Hickman group implemented such an approach in experiments with hippocampal neurons. 21 Further research has led to the successful adoption of defined and serum-free medium formulations for (i) single cultures of skeletal myotubes, 22 cardiomyocytes, 23 and motoneurons 20,24 ; (ii) dual cultures of motoneurons with skeletal muscle (to form neuromuscular junctions), 17 sensory neurons (to recreate the stretch reflex arc), 16,25 or oligodendrocyte progenitor cells (to model myelination) 26 ; and (iii) multi-organ cultures with four different organ types. 7…”